Rapid pathogen detection, contamination and bioburden monitoring, particularly for safety and quality control testing of foods, including beverages and drinking water, agricultural, including GMO-modified foods, environmental, including cell culture media, biopharmaceuticals, including the manufacturing process, and pharmaceuticals, including parenteral drugs, is an ongoing challenge. Microbial contaminants are a major concern in the biopharmaceutical industry and for regulatory organizations, including those charged with public health and safety. Contamination by microorganisms is therefore of significant concern to the biopharmaceutical industry, which is dependent on continuous cell culture for the production of drugs, vaccines, and other “biologics”. Likewise, when testing for the bioburden in pharmaceutical manufacturing of biopharmaceuticals, the nucleic acid from non-viable contaminants would still be detectable in an amplification reaction. Conversely, use of actual pathogen nucleic acid controls creates the potential for contamination of test samples and false positive results.
To address these concerns, described herein is a discriminating positive control that confirms nucleic acid extraction, amplification as well as providing differentiation of a true positive in a sample from a false positive due to inadvertent contamination of the test sample with positive control DNA. Further, the discriminating positive control will eliminate or reduce the concern of contamination resulting from supplementing a positive control reaction with nucleic acid derived from the microorganism of interest.